Abomasal dysfunction and cellular and mucin changes during infection of sheep with larval or adult Teladorsagia circumcincta.

This is the first integrated study of the effects on gastric secretion, inflammation and fundic mucins after infection with L3 T. circumcincta and in the very early period following transplantation of adult worms. At 3 months-of-age, 20 Coopworth lambs were infected intraruminally with 35,000 L3; infected animals were killed on Days 5, 10, 15, 20 and 30 post-infection and 6 controls on either Day 0 or 30 post-infection. Another 15 Romney cross lambs received 10,000 adult worms at 4-5 months-of-age though surgically-implanted abomasal cannulae and were killed after 6, 12, 24 and 72 hours; uninfected controls were also killed at 72 hours. Blood was collected at regular intervals from all animals for measurement of serum gastrin and pepsinogen and abomasal fluid for pH measurement from cannulated sheep. Tissues collected at necropsy were fixed in Bouin's fluid for light microscopy, immunocytochemistry and mucin staining and in Karnovsky's fluid for electron microscopy. Nodules around glands containing developing larvae were seen on Day 5 p.i., but generalised effects on secretion occurred only after parasite emergence and within hours after transplantation of adult worms. After L3 infection, there were maximum worm burdens on Days 10-15 post-infection, together with peak tissue eosinophilia, inhibition of gastric acid secretion, hypergastrinaemia, hyperpepsinogenaemia, loss of parietal cells, enlarged gastric pits containing less mucin and increased numbers of mucous neck cells. After adult transplantation, serum pepsinogen was significantly increased after 9 hours and serum gastrin after 18 hours. Parallel changes in host tissues and the numbers of parasites in the abomasal lumen suggest that luminal parasites, but not those in the tissues, are key drivers of the pathophysiology and inflammatory response in animals exposed to parasites for the first time. These results are consistent with initiation of the host response by parasite chemicals diffusing across the surface epithelium, possibly aided by components of ES products which increased permeability. Parietal cells appear to be a key target, resulting in secondary increases in serum gastrin, pit elongation, loss of surface mucins and inhibition of chief cell maturation. Inflammation occurs in parallel, and could either cause the pathology or exacerbate the direct effects of ES products.

Anti-parasitic activity of pelleted sainfoin (Onobrychis viciifolia) against Ostertagia ostertagi and Cooperia oncophora in calves.

BACKGROUND: Increasing anthelmintic-resistance in nematodes of ruminants emphasises the need for sustainable parasite control. Condensed tannin-containing legume forages such as sainfoin (Onobrychis viciifolia) have shown promising anthelmintic properties in small ruminants but this has never been explored in cattle. Therefore, our aim was to examine the efficacy of sainfoin against cattle nematodes in vivo. METHODS: Fifteen Jersey male calves (2-4 month-old) were allocated into two groups and fed isoproteic and isoenergetic diets mainly composed of sainfoin pellets (Group SF; n = 9, three pens) or concentrate and grass-clover hay (Group CO; n = 6, two pens). After 16 days of adaptation, all animals were experimentally infected with 10,000 and 66,000 third-stage larvae of Ostertagia ostertagi and Cooperia oncophora, respectively. Egg excretion, blood parameters and bodyweights were recorded throughout the study. Worms were harvested by sieving for quantification and scanning electron microscopy (SEM) 42 days post-infection (dpi) when the calves were necropsied. RESULTS: The number of O. ostertagi adults in the abomasum was reduced by 50 % in Group SF compared with Group CO (P < 0.05). This was further reflected in higher albumin (P < 0.1) and lower pepsinogen levels (P < 0.05) in Group SF at 21 dpi, and structural damage of the worm cuticle could be visualised by SEM. Yet, the nematode egg excretion in Group SF was not significantly different from that of the controls (P > 0.05). Likewise, no statistical difference in total worm burdens of C. oncophora was found between the groups. Weight gains were lower for Group SF (P < 0.05), which may reflect lower digestibility and phosphorus levels in the SF diet, despite similar feed intake at pen-level. CONCLUSIONS: Overall, the effect of sainfoin on abomasal nematodes corroborates results from studies with small ruminants and encourages further investigations of the use of this crop for control of cattle nematodes.

The Effect of Marshallagia marshalli on Serum Gastrin Concentrations in Experimentally Infected Lambs.

: Because there appeared to be no data available on serum gastrin concentrations in animals infected with Marshallagia marshalli, and considering the high prevalence of this parasite in livestock throughout many countries, we decided to perform research in the field using experimental infection. After surgical implantation of abomasal cannula into 10 male Baluchi sheep, each animal was orally infected with 5,000 M. marshalli larvae. Serum gastrin concentrations and abomasal pH were measured with a human ELISA kit and a PHM LE438 standard pH electrode, respectively. According to the results obtained from the study, serum gastrin increased after 14 and 21 days post-infection (dpi), while abomasal pH increased after 7 dpi and reached a maximal value 16 dpi. The increase in serum gastrin concentration was revealed 6 days after elevation in abomasal pH, which could be the result of reduced acid secretion. Generally, the present study pointed out that a limited number of M. marshalli could increase serum gastrin concentrations.

Faecal dry matter, inflammatory cells and antibodies in parasite-resistant sheep challenged with either Trichostrongylus colubriformis or Teladorsagia circumcincta.

Sheep that are highly resistant to parasitic nematodes can suffer bad diarrhoea due to the inflammation associated with rejection of ingested larvae from pasture. We hypothesised that challenging parasite-resistant sheep indoors with nematode larvae would result in reduced faecal dry matter, and that faecal dry matter would be lowest in sheep challenged with a mixture of Trichostrongylus colubriformis and Teladorsagia circumcincta compared to those challenged with either species alone. We also hypothesised that inflammatory cells and serum antibodies and interleukin-5 would be highest in those sheep that received the mixed larval challenge. We found that faecal dry matter was reduced (P<0.05) in challenged sheep compared to unchallenged sheep, with the fastest reduction being in those sheep challenged with only Tric. colubriformis. At 14 and 23 days after challenge began, there were no differences in faecal dry matter between the three challenged groups. Within the abomasum, there were no differences in inflammatory cell numbers between unchallenged sheep and those challenged only with Tric. colubriformis. Cell numbers in sheep challenged with Tela. circumcincta were higher (P<0.05) than those in unchallenged sheep, but there were no differences between sheep challenged only with Tela. circumcincta or as a mixed challenge. In the small intestine, inflammatory cell numbers were higher (P<0.05) in sheep that received the mixed challenge compared to controls. Cell numbers in sheep challenged with either Tela. circumcincta or Tric. colubriformis were also slightly higher than those in controls. Larval challenge increased (P<0.05) levels of IgA and IgE in serum, but there were no differences between the three challenged groups. Larval challenge also increased (P<0.05) levels of IL-5, with the greatest increase being in those sheep challenged with both species. We concluded that both Tela. circumcincta and Tric. colubriformis can cause immune-mediated diarrhoea in sheep, and that a mixed challenge will not necessarily lead to worse diarrhoea or higher concentrations of antibodies in serum. We also concluded that challenge with Tric. colubriformis leads to no inflammation in the abomasum, but challenge with Tela. circumcincta may lead to some inflammation in the small intestine.

[Some biochemical parameters in sheep infected with endoparasites (Fasciola spp., Dicrocoelium dendriticum, hydatid cysts, Trichostrongylidae and Protostrongylidae)]. / Endoparazitli (Fasciola spp., Dicrocoelium dendriticum, kist hidatik, Trichostrongylidae ve Protostrongylidae) koyunlarda bazi biyokimyasal parametreler.

This study was performed in order to investigate the variations of some blood biochemical parameters as well as the levels of Vitamin. B(12) and some macro elements in sheep infected with endoparasites. The blood samples were taken from the sheep that were to be slaughtered in the Van Municipality Slaughterhouse while the stool samples were taken after the slaughtering of the same animals. The postmortem examinations were made to investigate for the presence of Fasciola spp., D. dendriticum and cyst hydatid infections. The stool samples were examined helminthologically using native, sedimentation, flotation and Baermann-Wetzel methods. The control group was composed of animals not showing any internal organ parasites or parasites in the stool examination. Following the macroscopic and the stool examination, the animals found to have the same type of parasites were considered to be the study group. According to the analyses performed on the animals, the levels of total protein (in Trichostrongylidae, hydatid cysts), globulin, amylase, chlorine, and Vit.B(12) were found to be increased significantly, while the levels of albumin, magnesium, and phosphorus were found to be decreased significantly. The other parameters analyzed were not significant statistically between the groups.

Impact on productivity of peri-parturient rise in fecal egg counts in Creole goats in the humid tropics.

The control of gastrointestinal nematodes requires an understanding of their epidemiology so that particular parasite stages can be targeted. Dam infection during early lactation is one example of this in ruminant nematode infections. The existence of the peri-parturient relaxation in immunity and its impact on productivity were examined in a Creole goat flock from Guadeloupe, exposed to mixed natural infection (predominantly Haemonchus contortus and Trichostrongylus colubriformis). A total of 1,511 l were obtained from 909 does resulting from 463 dams and 150 sires. Fecal and blood samples were collected at kidding before anthelmintic drenching, 4 and 6 weeks after kidding. The traits analyzed were logarithm transformed fecal egg counts (FEC), packed cell volume (PCV), and logarithm transformed blood eosinophilia counts (EOS) for does at each sampling point and changes in these during the postpartum period. With the exception of the PCV values measured at kidding, lactating does had significantly higher FEC and lower PCV than control dry does at every sampling point. Geometric means of FEC in lactating does were 819 +/- 174, 677 +/- 146 and, 699 +/- 160 eggs per gram (EPG) at kidding, 4 and 6 weeks after kidding respectively. Geometric means of FEC in dry does were 187 +/- 57, 89 +/- 28, 133 +/- 43 at these time points, respectively. EOS differences were not consistent between groups and probably not specific enough for variations in Creole goats' peri-parturient rise to be discussed. As does aged, their egg output decreased and primiparous does always had greater egg output than multiparous ones. Overall, does' FEC at 4 weeks after kidding decreased by 1.3% each year. The higher the litter size, the higher the FEC at kidding and inverse applied for PCV measurements. Does that stopped lactating had significantly lower FEC and higher PCV values than lactating does with low milk yields. Higher infection rates during early lactation in Creole goats were recorded in does with lower maternal ability assessed by the average daily weight gain of kids between 10 and 30 days of age. Kids from dams with higher FEC (i.e. >600 EPG higher than corresponding does) had 17% lower average daily weight gain between 30 and 70 days postpartum and were approximately 1 kg lighter at weaning than kids from dams with lower FEC. Thus, it is clear that a peri-parturient rise in FEC exists in Creole goats. By controlling the intensity of this peri-parturient rise in FEC, herd health and productivity could be substantially improved.

Genetic resistance to Teladorsagia circumcincta: IgA and parameters at slaughter in Churra sheep.

SUMMARY Previous experiments have shown that genetic resistance to infection by Teladorsagia circumcincta in sheep can be measured by the level of IgA in gastric mucus, jointly with other parameters. The aim of this study has been to observe the influence of IgA on adult worms. The experiment was carried out with Churra sheep experimentally infected with T. circumcincta. At slaughter, gastric content, gastric mucus, blood samples and faeces were recovered to determine the number of eggs in utero, length of adult females, worm burden, number of L4, titre of serum pepsinogen, peripheral eosinophilia and eggs per gram (epg). IgA activity in gastric mucus, serum, nasal secretions and saliva were tested against somatic antigen from fourth-stage larvae (L4), somatic antigen from the adult stage and excretory-secretory (E/S) antigen from the adult stage. The results showed a significant correlation between serum IgA and gastric mucus (P<0.01) as well as in nasal secretions (P<0.01). We found negative correlations between IgA activity in gastric mucus with the eggs in utero and with adult female length. Furthermore there were also strong relationships between the peripheral eosinophilia with serum (P<0.01) and gastric mucus IgA activity (P<0.01). Moreover serum pepsinogen and the number of L4 at slaughter were related (P<0.01).

Survey of gastric lesions and blood pepsinogen levels in pigs in Burkina Faso.

The purpose of the study was to investigate the prevalence of gastric lesions and to provide diagnostic values for serum pepsinogen in non-infected pigs and in pigs with gastric disease. In an abattoir survey, the pepsinogen concentrations were measured in the serum from 62 non-infected pigs, 33 pigs with gastric lesions and 17 pigs infected with Hyostrongylus rubidus, using a specific radioimmunoassay (RIA). The mean (+/- SE) pepsinogen concentrations in the serum of non-infected pigs, in pigs with gastric ulcers, and in pigs with a heavy H. rubidus infection were 630.8 +/- 39.2 ng/ml, 1084.5 +/- 166.2 ng/ml and 1095.2 +/- 102.3 ng/ml, respectively (p<0.05). Because of the higher concentrations of pepsinogen in the blood of pigs with gastric ulcers or parasitic infections, it is suggested that the measurement of serum pepsinogen by RIA may be an effective biochemical approach to the diagnosis of chronic gastric disorders in pigs.

Fructosamine concentration and resistance to natural, predominantly Teladorsagia circumcincta infection.

Fructosamine concentrations reflect protein status and because infection with Teladorsagia circumcincta can induce a relative protein deficiency, we examined the usefulness of fructosamine concentrations as markers of the intensity of infection in naturally infected lambs. Fructosamine concentration was a heritable trait and variation in fructosamine concerntrations was associated with differences in body weight, and a variety of parasitological variables; animals with increased fructosamine concentrations grew more quickly, had increased faecal egg counts in one of the three study years, had decreased pepsinogen concentrations and decreased IgA activity against 4th-stage larvae of T. circumcincta. Fructosamine concentrations were also associated with variation in the subsequent acquisition of nematodes and in the length of adult female T. circumcincta; lambs with increased fructosamine concentrations had fewer nematodes but the mean length of adult female T. circumcincta was longer. Therefore fructosamine concentrations are potentially useful indicators of the severity of nematode infection and may predict magnitude of subsequent infection.

Studies on the immunoglobulin E responses to Teladorsagia circumcincta in sheep: purification of a major high molecular weight allergen.

Studies on the immunoglobulin (Ig)E immune responses to the gastric nematode, Teladorsagia circumcincta, have demonstrated a major high molecular weight allergen (HMWTc). Cross reactive allergens of similar MW were demonstrated for Trichostrongylus colubriformis and Cooperia curticei, but not for Haemonchus contortus. Purification of HMWTc was achieved by gel-filtration chromatography, and nonreducing SDS-PAGE and Western blot analysis revealed two closely associated bands with a molecular weight of approximately 140-150 kDa. Reduction showed four IgE reactive bands of 120, 50, 45 and 30 kDa, and deglycosylation abrogated the immunoreactivity of the 120 and 30 kDa bands. Ultrastructural immunolocalization by electron microscopy revealed that the IgE reactivity was confined to the cuticular surface of the infective (L3) larvae. ELISA studies to determine the IgE anti-HMWTc responses in lambs during their first grazing season, demonstrated significantly higher IgE antibody in lambs with low accumulative faecal egg count (FEC) compared to animals with high accumulative FEC. These studies provide evidence for a protective function of IgE antibody in Teladorsagia infections in lambs.

Comparative aspects of plasma antioxidant status in sheep and goats, and the influence of experimental abomasal nematode infection.

This paper provides, for the first time, comparative data on the plasma antioxidant status of two ruminant species, namely sheep and goats. In addition, the influence of experimental infection with Teladorsagia circumcincta on antioxidant status in the same two species is compared and contrasted. In general terms, antioxidant status was significantly higher in uninfected kids than in lambs. Differences in protein sulphydryl groups and vitamin E concentrations were particularly noteworthy; trends were similar, however, for albumin, vitamin A and total antioxidant capacity (TAC). Parasitological results, based on worm burden, faecal egg counts and peripheral blood eosinophil numbers, confirmed that goat kids were more susceptible than lambs to experimental T. circumcincta infection. "Trickle infection" had a variable impact on both total and individual antioxidant status; particularly during the early weeks, the trend was for reduced values in lambs and increased values in kids, as compared with uninfected controls. Subsequent challenge infection was associated with a transient decrease in TAC and albumin in trickle-infected animals of both species, and in appropriate control animals. The observed differences in plasma antioxidant capacity between sheep and goats may have important implications in terms of the comparative resilience of sheep and goats to parasite infection.

The mast cell and eosinophil response of young lambs to a primary infection with Nematodirus battus.

Helminthologically naive 8-week-old lambs were administered 50,000 infective-stage larvae of the trichostrongyle nematode Nematodirus battus. Initial mean adult worm burdens were significantly reduced (P < 0.01) at day 21 post-infection (p.i.). Peripheral blood eosinophil counts showed a significant increase (P < 0.001) by day 25 p.i. when compared with uninfected control animals. Sections of small intestine taken at post-mortem from the area of adult worm establishment over the course of infection showed a significant increase in mucosal eosinophil and mast cell numbers during the period of adult worm rejection, when compared with uninfected control animals. These results show that young lambs develop significant increases in the numbers of cells associated with an inflammatory reaction in the intestine during the development of a partially protective immune response to this parasitic nematode.

Haemopoietic cell responses in the blood and bone marrow of sheep infected with the abomasal nematode Telodorsagia circumcincta.

The generation of bone marrow and blood haemopoietic progenitor colony-forming cells (CFCs) in sheep given primary or challenge infections with the nematode parasite Telodorsagia circumcincta is described. Ten days after a primary infection, the frequency of early multipotential-CFCs, eosinophil-CFCs, macrophage-CFCs and mast cell or basophil-CFCs was greater than in controls. These frequencies then declined to pre-infection levels by day 21. Blood CFCs (mainly macrophage-CFCs and eosinophil-CFCs) also increased after infection, indicating a migration of CFCs, presumably to the site of infection. Ten days after challenge infection there was less marked myelopoiesis than in the primary infection on day 10, though both eosinophil-CFCs and mast cell or basophil-CFCs were significantly above control values. Blood CFC output (mainly macrophage-CFCs and eosinophil-CFCs) reached a peak 2-6 days after challenge, evidence of rapid recruitment to the site of infection. Telodorsagia circumcincta infection is therefore associated with an increase in myelopoiesis, particularly for the cell types characteristic of the local inflammatory response to abomasal nematodes. There was no correlation between any of the haemopoietic cell responses measured and worm burdens in individual animals after either primary or challenge infection.

Primary infection of kids with Teladorsagia circumcincta: susceptibility and blood constituents.

Fourteen male cross-bred Alpine x Saanen kids were infected with Teladorsagia circumcincta and four were used as controls. Three different levels of susceptibility to infection were apparent: resistant to infection and resilient to its adverse effects; intermediate (poorly resistant and poorly resilient); susceptible and relatively resilient. In the susceptible kids the establishment rate of infective larvae was twice as large and the nematode egg output five times greater during Weeks 4-8 after infection than in resistant kids. Susceptible kids harboured larger females with higher numbers of eggs in utero; the proportion of Teladorsagia trifurcata was higher in susceptible than in resistant kids. The blood constituents were significantly modified in infected compared with control animals: a lower number of leucocytes (particularly lymphocytes and basophils) and a higher complement activity were recorded in the latter. Compared with resistant kids, susceptible kids had higher counts of leucocytes, basophils and lower lymphocyte levels, less depleted complement pathway activity, higher pepsinogen values and lower levels of albumin and glucose.

Levels of serum IgE in response to gastrointestinal nematodes in cattle.

Serum IgE concentration has been considered a valuable measurement in parasitic infections, yet little has been reported regarding cattle. This study examines the association of IgE levels of nematode-naïve Holstein steer calves and the level of gastrointestinal parasitism acquired by grazing irrigated pasture for 30 days. Total IgE levels were determined by enzyme-linked immunosorbent assay (ELISA) specific for bovine IgE on serum collected both before and after exposure to parasite-infected pastures. Following necropsy, parasite loads were determined by direct count from the contents of the abomasum, small intestine, cecum and large intestine; species of Ostertagia and Cooperia being the most common helminths found. Significant increases of IgE in the serum of calves with light infestations were seen, whereas calves with moderate to heavy infestations showed only mild IgE increases. With increased parasite burden, the frequency of increased IgE levels was reduced. Additionally, there appeared to be a seasonal correlation relating the level of serum IgE detected to the number of worms counted and to the course of parasite development.

Pathophysiology of gastrointestinal trichostrongyles in sheep: plasma losses and changes in plasma pepsinogen levels associated with parasite challenge of immune animals.

The pathophysiological effects of experimental larval challenge with a mixed trichostrongyle infection were measured in ewes which had previously grazed contaminated pasture. Their immune status was confirmed by low worm burdens, both before and after challenge. Despite the low worm burdens, it was evident that larval challenge was associated with marked pathophysiological disturbances and, in particular, a doubling of plasma pepsinogen levels, albumin catabolism and losses of plasma protein into the gastrointestinal tract. The concurrent timing of these changes suggested they had a common aetiology. These findings indicate that larval challenge of such animals may cause impaired production, despite low parasite burdens.

Plasma protein loss associated with gastrointestinal parasitism in grazing sheep.

Some pathophysiological effects of parasitic gastroenteritis in two groups of lambs grazing paddocks either heavily or lightly contaminated with trichostrongyle larvae were investigated between July and October 1980. The leak of plasma protein was measured on three occasions at pasture using 51chromic chloride. Total faecal output was measured indirectly using chromic oxide. Losses of 51chromic chloride-labelled plasma protein into the gastrointestinal tract were significantly higher in the lambs grazing the heavily contaminated pasture than in those grazing lightly infected ground in both July and August. The increased plasma losses were associated with high faecal egg counts, hypoalbuminaemia and elevated levels of plasma pepsinogen.

Experimental infection of Nematospiroides dubius in Swiss albino mice. IX. Effect of dose break up on the serum protein patterns.

The present study concerns with the effects of larval inoculation with either a single dose or two or more spaced doses of 200 larvae on the serum protein profiles. No significant difference occurred in the various fractions among the three groups indicating that the total quantity of antigenic stimulation in the most important factor in eliciting such changes, no matter whether the dose is given once or spaced in two or three small doses. Serum protein fractions of Swiss albino mice undergoing experimental infection of Nematospiroides dubius do not tend to rise further due to repeated exposures in comparison to those that received a challenge dose only (1). Hence, experiments were designed to ascertain whether there is any difference in serum protein patterns due to breaking up of the infective dose into two or three spaced doses of N. dubius larvae.